改良植块法培养原代成骨细胞

目的:应用改良植块法体外培养大鼠颅骨成骨细胞.方法:将1～3天的新生SD大鼠的颅骨剪碎、接种于培养瓶底,当成骨细胞从组织块中爬出、融合成单层,消化收集成骨细胞,组织块则继续进行培养.将分批次得到的成骨细胞进行MTT,ALP活性以及钙结节计数分析.结果:利用该方法连续分次培养出的成骨细胞.经形态学及ALP鉴定符合成骨细胞的生物学特性,MTT,ALP活性以及钙结节计数无统计学差异.结论:连续植块法培养成骨细胞,操作简便,成功率高,能持续提供细胞,满足实验需求.

Osteoblastic cell cultured with modified explant culture

Objective:To culture rat cranial osteoblastic cells in vitro by modified explants culture.Methods:Primary osteoblastic cells were obtained from the calvaria of 3-day-old new-born SD rats.The calvaria were divided into small pieces and seeded on bottom of culture bottle.When osteoblasts crawled from the explants and confluented to monolayer,they were digested and aggregated by 0.25% trypsin and explants continued culturing.MTT,ALP activity and calcific nodule were analysized in different osteoblasts.Results:The cultured cells by this method were identified according to the osteoblastic bionomics.There were no statistical differences in MTT,ALP activity and calcific nodule among the osteoblasts.Conclusion:The modified explants method to culture primary osteoblasts could obtain large amount of cells and supply osteoblasts continuously.